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Cell Cycle: The complex series of phenomena, occurring between the end of one Cell division and the end of the next, by which cellular material is duplicated and then divided between two daughter cells. The cell cycle includes Interphase, which includes G0 phase; G1 phase; S Phase; and G2 phase, and Cell division phase.

What is the Cell Cycle?

JoVE 10757

The cell cycle refers to the sequence of events occurring throughout a typical cell’s life. In eukaryotic cells, the somatic cell cycle has two stages: interphase and the mitotic phase. During interphase, the cell grows, performs its basic metabolic functions, copies its DNA, and prepares for mitotic cell division. Then, during mitosis and cytokinesis, the cell divides its nuclear and cytoplasmic materials, respectively. This generates two daughter cells that are identical to the original parent cell. The cell cycle is essential for the growth of the organism, replacement of damaged cells, and regeneration of aged cells. Cancer is the result of uncontrolled cell division sparked by a gene mutation. There are three major checkpoints in the eukaryotic cell cycle. At each checkpoint, the progression to the next cell cycle stage can be halted until conditions are more favorable. The G1 checkpoint is the first of these, where a cell’s size, energy, nutrients, DNA quality, and other external factors are evaluated. If the cell is deemed inadequate, it does not continue to the S phase of interphase. The G2 checkpoint is the second checkpoint. Here, the cell ensures that all of the DNA has been replicated and is not damaged before entering mitosis. If any DNA damage is detected that cannot be repaired, the cell may undergo apoptosis, or

 Core: Cell Cycle and Division

Cell Cycle Analysis

JoVE 5641

Cell cycle refers to the set of events through which a cell grows, replicates its genome, and ultimately divides into two daughter cells through the process of mitosis. Because the amount of DNA in a cell shows characteristic changes throughout the cycle, techniques known as cell cycle analysis can be used to separate a population of cells according to the different phases …

 Cell Biology

Cell Division- Concept

JoVE 10571

Cell division is fundamental to all living organisms and required for growth and development. As an essential means of reproduction for all living things, cell division allows organisms to transfer their genetic material to their offspring. For a unicellular organism, cellular division generates a completely new organism. For multicellular organisms, cellular division produces new cells for…

 Lab Bio

Positive Regulator Molecules

JoVE 10763

To consistently produce healthy cells, the cell cycle—the process that generates daughter cells—must be precisely regulated.

Internal regulatory checkpoints ensure that a cell’s size, energy reserves, and DNA quality and completeness are sufficient to advance through the cell cycle. At these checkpoints, positive and negative regulators promote or inhibit a cell’s continuation through the cell cycle. Positive regulators include two protein groups that allow cells to pass through regulatory checkpoints: cyclins and cyclin-dependent kinases (CDKs). These proteins are present in eukaryotes, ranging from yeast to humans. Cyclins can be categorized as G1, G1/S, S, or M cyclins based on the cell cycle phase or transition they are most involved in. Generally, levels of a given cyclin are low during most of the cell cycle but abruptly increase at the checkpoint they most contribute to (G1 cyclins are an exception, as they are required throughout the cell cycle). The cyclin is then degraded by enzymes in the cytoplasm and its levels decline. Meanwhile, cyclins needed for the next checkpoints accumulate. To regulate the cell cycle, cyclins must be bound to a Cyclin-dependent kinase (CDK)—a type of enzyme that attaches a phosphate group to modify the activity of a target protein.

 Core: Cell Cycle and Division

Cell Division - Student Protocol

JoVE 10572

Observing the Cell Cycle in a Root Tip
Hypotheses: The experimental hypothesis is that in root tips slices that have been treated with nocodazole, a chemical that interferes with microtubular polymerization, all of the cells will be arrested at the same stage of the cell cycle and that in untreated onion tip slices all of the different stages of the…

 Lab Bio

Negative Regulator Molecules

JoVE 10764

Positive regulators allow a cell to advance through cell cycle checkpoints. Negative regulators have an equally important role as they terminate a cell’s progression through the cell cycle—or pause it—until the cell meets specific criteria.

Three of the best-understood negative regulators are p53, p21, and retinoblastoma protein (Rb). The regulatory roles of each of these proteins were discovered after faulty copies were found in cells with uncontrolled replication (i.e., cancer). These proteins exert most of their regulatory effects at the G1 checkpoint early in the cell cycle. P53 strongly influences a cell’s commitment to divide. It responds to DNA damage by discontinuing the cell cycle and summoning enzymes to repair the damage. If the DNA damage is irreparable, p53 can prevent the cell from proceeding through the cell cycle by inducing apoptosis, or cell death. An increase in p53 triggers the production of p21. P21 prevents the cell from transitioning from the G1 to the S phase of the cell cycle by binding to CDK/cyclin complexes, inhibiting their positive regulatory actions. Rb negatively regulates the cell cycle by acting on different positive regulators, mainly in response to cell size. Active (dephosphorylated) Rb binds to transcription factors, preventing them from initiating gene tran

 Core: Cell Cycle and Division

Magnetic Activated Cell Sorting (MACS): Isolation of Thymic T Lymphocytes

JoVE 10495

Source: Meunier Sylvain1,2,3, Perchet Thibaut1,2,3, Sophie Novault4, Rachel Golub1,2,3
1 Unit for Lymphopoiesis, Department of Immunology, Pasteur Institute, Paris, France
2 INSERM U1223, Paris, France
3 Université Paris Diderot, Sorbonne Paris Cité, Cellule Pasteur, Paris, France
4 Flow Cytometry Platfrom, Cytometry and Biomarkers UtechS, …

 Immunology

Mitosis and Cytokinesis

JoVE 10762

In eukaryotic cells, the cell's cycle—the division cycle—is divided into distinct, coordinated cellular processes that include cell growth, DNA replication/chromosome duplication, chromosome distribution to daughter cells, and finally, cell division. The cell cycle is tightly regulated by its regulatory systems as well as extracellular signals that affect cell proliferation. The processes of the cell cycle occur over approximately 24 hours (in typical human cells) and in two major distinguishable stages. The first stage is DNA replication, during the S phase of interphase. The second stage is the mitotic (M) phase, which involves the separation of the duplicated chromosomes into two new nuclei (mitosis) and cytoplasmic division (cytokinesis). The two phases are separated by intervals (G1 and G2 gaps), during which the cell prepares for replication and division. Mitosis can be divided into five distinct stages—prophase, prometaphase, metaphase, anaphase, and telophase. Cytokinesis, which begins during anaphase or telophase (depending on the cell), is part of the M phase, but not part of mitosis. As the cell enters mitosis, its replicated chromosomes begin to condense and become visible as threadlike structures with the aid of proteins known as condensins. The mitotic spindle apparatus b

 Core: Cell Cycle and Division

An Introduction to Cell Division

JoVE 5640

Cell division is the process by which a parent cell divides and gives rise to two or more daughter cells. It is a means of reproduction for single-cell organisms. In multicellular organisms, cell division contributes to growth, development, repair, and the generation of reproductive cells (sperms and eggs). Cell division is a tightly regulated process, and aberrant cell…

 Cell Biology

Cell Division - Prep Student

JoVE 10626

Preparation of Onion Root Tips and Solutions
For the cell cycle experiment using root tips, first, leave an onion suspended over a beaker of water to grow roots for several days.
Then, clean the onion roots of any dirt or debris.
Next, in a 1.5 mL tube, dissolve 10 mcg of nocodazole per 1 mL of dimethyl sulfoxide solution, making 1 tube…

 Lab Bio

Live Cell Imaging of Mitosis

JoVE 5642

Mitosis is a form of cell division in which a cell’s genetic material is divided equally between two daughter cells. Mitosis can be broken down into six phases, during each of which the cell’s components, such as its chromosomes, show visually distinct characteristics. Advances in fluorescence live cell imaging have allowed scientists to study this process in…

 Cell Biology

Interphase

JoVE 10761

The cell cycle occurs over approximately 24 hours (in a typical human cell) and in two distinct stages: interphase, which includes three phases of the cell cycle (G1, S, and G2), and mitosis (M). During interphase, which takes up about 95 percent of the duration of the eukaryotic cell cycle, cells grow and replicate their DNA in preparation for mitosis.

Following each period of mitosis and cytokinesis, eukaryotic cells enter interphase, during which they grow and replicate their DNA in preparation for the next mitotic division. During the G1 (gap 1) phase, cells grow continuously and prepare for DNA replication. During this phase, cells are metabolically active and copy essential organelles and biochemical molecules, such as proteins. In the subsequent S (synthesis) phase of interphase, cells duplicate their nuclear DNA, which remains packaged in semi-condensed chromatin. During the S phase, cells also duplicate the centrosome, a microtubule-organizing structure that forms the mitotic spindle apparatus. The mitotic spindle separates chromosomes during mitosis. In the G2 (gap 2) phase, which follows DNA synthesis, cells continue to grow and synthesize proteins and organelles to prepare for mitosis. In human cells, the G1 phase spans approximately 11 hours, the S phase takes about

 Core: Cell Cycle and Division

Yeast Reproduction

JoVE 5097

Saccharomyces cerevisiae is a species of yeast that is an extremely valuable model organism. Importantly, S. cerevisiae is a unicellular eukaryote that undergoes many of the same biological processes as humans. This video provides an introduction to the yeast cell cycle, and explains how S. cerevisiae reproduces both asexually and sexually Yeast reproduce asexually …

 Biology I

An Introduction to Saccharomyces cerevisiae

JoVE 5081

Saccharomyces cerevisiae (commonly known as baker’s yeast) is a single-celled eukaryote that is frequently used in scientific research. S. cerevisiae is an attractive model organism due to the fact that its genome has been sequenced, its genetics are easily manipulated, and it is very easy to maintain in the lab. Because many yeast proteins are similar in sequence and function…

 Biology I

Yeast Maintenance

JoVE 5095

Research performed in the yeast Saccharomyces cerevisiae has significantly improved our understanding of important cellular phenomona such as regulation of the cell cycle, aging, and cell death. The many benefits of working with S. cerevisiae include the facts that they are inexpensive to grow in the lab and that many ready-to-use strains are now commercially available. Nevertheless,…

 Biology I

The Nucleus

JoVE 10691

The nucleus is a membrane-bound organelle that contains a eukaryotic organism’s genetic instructions in the form of chromosomal DNA. This is distinct from the DNA in mitochondria or chloroplasts that carry out functions specific to those organelles. While some cells—such as red blood cells—do not have a nucleus, and others—such as skeletal muscle cells—have multiple nuclei, most eukaryotic cells have a single nucleus. The DNA in the nucleus is wrapped around proteins such as histones, creating a DNA-protein complex called chromatin. When cells are not dividing—that is, when they are in the interphase part of their cell cycle—the chromatin is organized diffusely. This allows easy access to the DNA during the transcription process when messenger RNA (mRNA) is synthesized based on the DNA code. When a eukaryotic cell is about to divide, the chromatin condenses tightly into distinct, linear chromosomes. Humans have 46 chromosomes in total. Chromatin is particularly concentrated in a region of the nucleus called the nucleolus. The nucleolus is important for the production of ribosomes, which translate mRNA into protein. In the nucleolus, ribosomal RNA is synthesized and combined with proteins to create ribosomal subunits, which later form functioning ribosomes in the cytoplasm of the cell. The interior of t

 Core: Cell Structure and Function

Meiosis II

JoVE 10768

Meiosis II is the second and final stage of meiosis. It relies on the haploid cells produced during meiosis I, each of which contain only 23 chromosomes—one from each homologous initial pair. Importantly, each chromosome in these cells is composed of two joined copies, and when these cells enter meiosis II, the goal is to separate such sister chromatids using the same microtubule-based network employed in other division processes. The result of meiosis II is two haploid cells, each containing only one copy of all 23 chromosomes. Depending on whether the process occurs in males or females, these cells may form eggs or sperm, which—when joined through the process of fertilization—may yield a new diploid individual. Although the goal of meiosis II is the same in both males and females—to produce haploid egg or sperm cells—there are some critical differences in this process between the sexes. For example, in a woman’s egg precursor cells, the meiotic spindle apparatus responsible for separating sister chromatids forms off to one side, near the periphery. This asymmetry allows for two cells of unequal sizes to be produced following meiosis II: a large egg, and a smaller polar body that dissolves. This division of cytoplasm ensures that the egg contains enough nutrients to support an embryo. The position of the meiotic spind

 Core: Meiosis

Passaging Cells

JoVE 5052

Cell lines are frequently used in biomedical experiments, as they allow rapid culture and expansion of cell types for experimental analysis. Cell lines are cultured under similar conditions when compared to freshly-isolated, or primary, cells, but with some basic important differences: (i) cell lines require their own specific growth factor cocktails and (ii) their growth must be more closely…

 Basic Methods in Cellular and Molecular Biology

Epigenetic Regulation

JoVE 10803

Epigenetic mechanisms play an essential role in healthy development. Conversely, precisely regulated epigenetic mechanisms are disrupted in diseases like cancer.

In most mammals, females have two X chromosomes (XX) while males have an X and a Y chromosome (XY). The X chromosome contains significantly more genes than the Y chromosome. Therefore, to prevent an excess of X chromosome-linked gene expression in females, one of the two X chromosomes is randomly silenced during early development. This process, called X-chromosome inactivation, is regulated by DNA methylation. Scientists have found greater DNA methylation at gene promoter sites on the inactive X chromosome than its active counterpart. DNA methylation prevents the transcription machinery from attaching to the promoter region, thus inhibiting gene transcription. Abnormal DNA methylation plays an important role in cancer. The promoter region of most genes contains stretches of cytosine and guanine nucleotides linked by a phosphate group. These regions are called CpG islands. In healthy cells, CpG islands are not methylated. However, in cancer cells, CpG islands in the promoter regions of tumor suppressor genes or cell cycle regulators are excessively methylated. Methylation turns off the expression of these genes, allowing cancer cells to divide rapidly and uncontrollably.

 Core: Gene Expression

Cancer

JoVE 10987

Cancers arise due to mutations in genes involved in the regulation of cell division, which leads to unrestricted cell proliferation. Modern science and medicine have made great strides in the understanding and treatment of cancer, including eradicating cancer in some patients. However, there is still no cure for cancer. This is largely due to the fact that cancer is a large group of many diseases. Tumors may result in a case where two people have the same mutations in an oncogene or tumor suppressor gene. Initially, the tumors may be very similar. However, the uncontrolled cell division results in new random mutations. As the tumor cells continue to divide, they become more varied. As a result, the two tumors will grow at different rates and undergo angiogenesis and metastasis at different times. The two cancers become so distinct from one another that they will not respond in the same way to the same therapy. This demonstrates why even a particular type of cancer, breast cancer, for example, can be a myriad of different cancers, each disease case with its unique properties, potentially requiring unique treatment approaches. As such, new cancer research and clinical trials focus on tailoring therapeutic approaches specifically for each patient’s genomic and molecular landscape. This is called personalized medicine. On the other hand, chemotherapy a

 Core: Cell Cycle and Division

Zebrafish Reproduction and Development

JoVE 5151

The zebrafish (Danio rerio) has become a popular model for studying genetics and developmental biology. The transparency of these animals at early developmental stages permits the direct visualization of tissue morphogenesis at the cellular level. Furthermore, zebrafish are amenable to genetic manipulation, allowing researchers to determine the effect of gene expression on the…

 Biology II

Nondisjunction

JoVE 11013

During meiosis, chromosomes occasionally separate improperly. This occurs due to failure of homologous chromosome separation during meiosis I or failed sister chromatid separation during meiosis II. In some species, notably plants, nondisjunction can result in an organism with an entire additional set of chromosomes, which is called polyploidy. In humans, nondisjunction can occur during male or female gametogenesis and the resulting gametes possess one too many or one too few chromosomes. When an abnormal gamete fuses with a normal gamete, the resulting zygote has an abnormal number of chromosomes and is called aneuploid. An individual with one too few chromosomes has monosomy (45; 2n-1), while trisomy is the presence of one too many chromosomes for a total of 47 (2n+1). Down Syndrome is one well-studied trisomy, where individuals have three copies of chromosome 21. Aneuploid zygotes account for around 70% of spontaneous abortions during gestation. Nondisjunction is more common in sex chromosomes than autosomes. Individuals can have a variety of sex chromosome combinations, including one or more additional sex chromosomes (e.g., XXY, XXX, XYY) or the presence of only a single sex chromosome (denoted X0). These individuals tend to have normal lifespans, though with sometimes major physiological and reproductive consequences. Nondisjunction appears to be more

 Core: Meiosis

Imaging- and Flow Cytometry-based Analysis of Cell Position and the Cell Cycle in 3D Melanoma Spheroids

1The Centenary Institute, 2Sydney Medical School, University of Sydney, 3The University of Queensland Diamantina Institute, Translational Research Institute, The University of Queensland, 4Department of Dermatology, Royal Prince Alfred Hospital, 5Discipline of Dermatology, University of Sydney

JoVE 53486

 Medicine

Analysis of Combinatorial miRNA Treatments to Regulate Cell Cycle and Angiogenesis

1School of Basic Pharmaceutical and Toxicological Sciences, College of Pharmacy, University of Louisiana Monroe, 2Department of Pharmaceutical Sciences, School of Pharmacy, Texas Tech University Health Sciences Center, 3Department of Obstetrics and Gynecology, School of Medicine, Texas Tech University Health Sciences Center

JoVE 59460

 Cancer Research

Live Cell Imaging and 3D Analysis of Angiotensin Receptor Type 1a Trafficking in Transfected Human Embryonic Kidney Cells Using Confocal Microscopy

1Department of Biochemistry, Georgetown University Medical Center, 2Department of Medicine, Georgetown University Medical Center, 3Department of Physics, Georgetown University Medical Center, 4Department of Oncology, Georgetown University Medical Center

JoVE 55177

 Biology

Mammalian Cell Division in 3D Matrices via Quantitative Confocal Reflection Microscopy

1Department of Chemical and Biomolecular Engineering, Johns Hopkins University, 2Johns Hopkins Physical Sciences - Oncology Center, Johns Hopkins University, 3Department of Biomedical Engineering, Johns Hopkins University, 4Departments of Oncology and Pathology and Sidney Kimmel Comprehensive Cancer Center, Johns Hopkins University School of Medicine

JoVE 56364

 Bioengineering

Stable and Efficient Genetic Modification of Cells in the Adult Mouse V-SVZ for the Analysis of Neural Stem Cell Autonomous and Non-autonomous Effects

1Cell Division and Cancer Group, Spanish National Cancer Research Centre (CNIO), 2Centro de Investigaciones Biomédicas en Red sobre Enfermedades Neurodegenerativas (CIBERNED), 3Departmento de Biologìa Celular, Universidad de Valencia, 4Institut de Biomedicina de la Universitat de Barcelona (IBUB), 5Department of Molecular and Translational Medicine, Fibroblast Reprogramming Unit, University of Brescia

JoVE 53282

 Developmental Biology

Identification of Nucleolar Factors During HIV-1 Replication Through Rev Immunoprecipitation and Mass Spectrometry

1Molecular and Cellular Biology Department, Beckman Research Institute at the City of Hope, 2Irell & Manella Graduate School of Biological Sciences, 3Department of Molecular Immunology, Beckman Research Institute and the City of Hope

JoVE 59329

 Immunology and Infection
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